Comparison of serological and molecular typing for HLA-A and -B on cord blood lymphocytes.
HLA class I typing by standard microcytotoxicity testing has been unsatisfactory for 14.5% of 1644 cord blood samples. In this study, we evaluated the capacity of PCR-SSP in solving problems in HLA-A,B typing with serological methods. With this aim we have compared serology with PCR-SSP in 100 cord blood samples with doubtful or unreliable HLA-A,B typing. PCR-SSP was successful in amplifying HLA-A,B alleles in all 100 cord blood samples. Forty-six typings gave discrepant results with the 2 methods (serology and PCR-SSP). Typings were considered discrepant also in the case of inability to define a split. For 19 specimens, no serological conclusion was drawn due to high mortality of the cell suspension, while PCR-SSP allowed the definition of a clear typing. In 6 cases it was necessary to infer information from serology to define the current typing. Finally, in 3 other cases it was impossible to exclude or attribute the antigen/allele B67 or B4802. PCR-SSP for HLA-A,B can improve the overall reliability of HLA-A,B typing requiring a small amount of blood although, with the set of sequence specific primers adopted, a number of alleles are still poorly defined.
Poli
F
F
Centro Trasfusionale e di Immunologia dei Trapianti, Ospedale Maggiore Policlinico - Istituto di Ricovero e Cura a Carattere Scientifico, Milano, Italy. nitpmi@mbox.vol.it
Scalamogna
M
M
Crespiatico
L
L
Macchi
B
B
Mistò
R
R
Nocco
A
A
Rossini
G
G
Scarpino
C
C
Sioli
V
V
Sirchia
G
G
D000483:Alleles; D005312:Fetal Blood / Q000166:cytology*; D015234:HLA-A Antigens / Q000145:classification / Q000235:genetics*; D015235:HLA-B Antigens / Q000145:classification / Q000235:genetics*; D006650:Histocompatibility Testing; D006801:Humans; D016133:Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000276:immunology*
D015234:HLA-A Antigens; D015235:HLA-B Antigens